Thermochimica Acta 457 (2007) 16

Compatibility studies of lapachol with pharmaceutical excipients

for the development of topical formulations
A.M. Lira
a
, A.A.S. Ara ujo
b
, I.D.J. Baslio
c
, B.L.L. Santos
a
,
D.P. Santana
a,
, R.O. Macedo
c
a
Departamento de Ci encias Farmac euticas, N ucleo de Desenvolvimento Farmac eutico e Cosm eticosNUDFAC,
Universidade Federal da Pernambuco, Recife, PE, CEP 50740-521, Brazil
b
Departamento de Fisiologia da Universidade Federal de Sergipe, Aracaju, SE, CEP 49000-000, Brazil
c
Laborat orio de Tecnologia Farmac eutica da Universidade Federal da Paraba,
Campus I Jo ao Pessoa, PB, CEP 58059-900, Brazil
Received 20 January 2006; received in revised form 14 February 2007; accepted 20 February 2007
Available online 27 February 2007
Abstract
Among naphthoquinones, lapachol and many heterocyclic derivatives have been investigated during the past years, mainly due to their antibac-
terial, antifungal and anticancer activities. Assessment of possible incompatibility between an active component (i.e. lapachol) and different
excipients along with the evaluation of thermal stability are crucial parts of a normal study prior to the nal formulation setting of a medicine.
DSC study was used as an important and complementary tool during pre-formulation to determine the compatibility of drug-excipients with the
purpose of developing a lapachol gel-cream formulation. The DSC curves of lapachol (Lpch) and binary mixtures with excipients (propylparaben,
methylparaben, edetic acid, isodecyl oleate, trietanolamine, glyceril monoesterate, cetiol, glicerin, carbopol, cetomacrogol and mineral oil) were
obtained. The results showed that Lpch only exhibited interaction which could inuence the stability of the product in the binary mixtures of
Lpch/methylparaben, Lpch/cetostearyl alcohol and Lpch/glyceryl monostearate. There was a signicant shift or absence of lapachol melting peak
in both cases. Photovisual DSC was used to conrm the results obtained with conventional DSC because it is a more sensitive method.
2007 Elsevier B.V. All rights reserved.
Keywords: Lapachol; DSC; Gel-cream; Compatibility studies
1. Introduction
Lapachol is a naphthoquinone (2-hydroxy-3-(3-methyl-
2-butenyl)-1,4-naphthoquinone) extracted from Pau darco
(Family Bignoneaceae). A great spectrum of therapeutic activ-
ities has been attributed to lapachol or its derivatives, such
as prevention of cercarial skin penetration of Schistossoma
mansoni [1,2], trypanosomicidal [3], antiinammatory [4] and
antineoplasic activity; antimalarial against erytrocytic stages of
Plasmodium falciparum [5] and against enteroviruses [68].
Assessment of possible incompatibility between an active
component (i.e. lapachol (Lpch)) and different excipients along
with the evaluation of thermal stability are crucial parts of a
normal study prior to the nal formulation setting of a topical

Corresponding author. Tel.: +55 79 32226614; fax: +55 79 32126640.

E-mail address: adriasa2001@yahoo.com.br (D.P. Santana).
dosage form [9]. Excipients are known to facilitate the admin-
istration and release of active components as well as to protect
them from the environment. Excipients are considered pharma-
ceutically inert, but physical and chemical interactions with an
active component are possible [9].
The development of pharmaceutical formulations requires
previous knowledge of the physicochemical properties of the
drug, excipients and analytical instrumentation that can be
applied efciently with swift results. Thermal analysis is
used in the pharmaceutical industry as a rapid technique
that is appropriate for quality control and the develop-
ment of new medicines [1012]. In particular, differential
scanning calorimetry (DSC) allows for the evaluation of pos-
sible incompatibilities by revealing changes in appearance,
shift or disappearance of melting or other exothermic pro-
cesses, and/or variations in the corresponding enthalpies of
reactions [13,14]. However, differences in the DSC curves
of binary mixtures compared to the individual components
0040-6031/$ see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.tca.2007.02.017
2 A.M. Lira et al. / Thermochimica Acta 457 (2007) 16
Fig. 1. DSC and TG/DTG curves of lapachol obtained in dynamic nitrogen
atmosphere (50 ml min
1
) and at a heating rate of 10

Cmin
1
.
may arise for reasons other than chemical incompatibility
[9].
This study evaluated the thermal stability of lapachol (Lpch)
and mixtures of Lpch/excipients by conventional DSC and pho-
tovisual system, andexcipients usedinthe prototype formulation
of gel-creams were selected. Gel-creams, which are relatively
new preparations, are oilwater systems containing gum as a
thickener in the external aqueous phase. They can be made from
a gelied preparation in which an emulsifying agent is incorpo-
rated in conjunction with an oily phase, which can be a mineral
or vegetable oil [15]. The use of a thickener increases the viscos-
ity of the continuous phase, reducing the frequency of collision
and coalescence of drops, thereby increasing the stability of the
preparation. Carbopol is commonly used as a thickener in these
types of formulations as its structure contains a small lipophilic
in addition to a large hydrophilic portion, which act as oil in
water emulsiers. The lipophilic part of this polymer is located
on the oilwater interface, and the hydrophilic portion swells in
the presence of water to formthe gel that envelops the tiny drops
of oil. This way, it is possible to obtain more stable formulations
that do not require a large quantity of oil phase to achieve a good
level of viscosity.
2. Experimental
2.1. Materials
Lapachol was donated by Laborat orio Farmac eutico do
Estado de Pernambuco (LAFEPE). Propylparaben, methyl-
paraben, edetic acid and isodecyl oleate were purchased from
Henrifarma. Trietanolamine and ethanol were obtained from
Merck. Glycerilmonoesterate was provided by Croda, and the
other excipients were acquired fromGalena. The mixed samples
consisted of equal weights of lapachol, and each excipient was
individually weighed into amber glass vials to give composite
weights of 20 mg. The physical mixtures were prepared using
an agata mortar with pestle for approximately 10 min.
2.2. Measurements
The DSC curves of the lapachol drug, excipients and binary
mixtures were obtained with a Shimadzu calorimeter, model
DSC-50, coupled to a photovisual system consisting of an
Olympus microscope connectedtoa Sanyocamera, model VCC-
D520, under a nitrogen ow of 50 ml min
1
, at a heating rate of
10

Cmin
1
, up to 400

C. These parameters were based on the

conventional laboratory programand appropriate method. Tran-
sition temperatures were recorded froma plot of heat owversus
temperature (30400

C). The peak temperature (T

p
) of phase
transitions was determined from the DSC curve with Tasys soft-
ware from Shimadzu. Reaction heat was determined by using
the area of the peaks between the onset temperature (T
o
) and the
end-temperature from the DSC curve. Samples (2.0 mg) were
weighed to the nearest 0.01 mg, and sealed aluminum cru-
cibles were used. The images were captured by means of DSC
coupled to the photovisual system under similar conditions of
conventional DSC. Mediumandstandarddeviation(S.D.) values
were determined from triplicates of DSC curves. The DSC cell
was calibrated with indium (mp 156.6

C; H
fus.
=28.5 J g
1
)
and zinc (mp 419.6

C). TG/DTG curves were obtained with a

thermobalance model TGA 50 (Shimadzu) in the temperature
Fig. 2. DSC photovisual of lapachol: (a) room temperature, (b) 138

C, (c) 139

C, (d) 140

C, (e) 183

C and (f) 196

C.
A.M. Lira et al. / Thermochimica Acta 457 (2007) 16 3
Fig. 3. DSCcurves of lapachol andsolidexcipients obtainedindynamic nitrogen
atmosphere (50 ml min
1
) and at a heating rate of 10

Cmin
1
.
range 25900

C, using platinum crucibles with 3 mg of sam-

ples, under dynamic nitrogen atmosphere (50 ml min
1
) and at
a heating rate of 10

Cmin
1
.
3. Results and discussion
DSC curves of lapachol showed a sharp endothermic
peak that corresponded to melting of Lpch at a temperature
Fig. 4. DSC curves of binary mixtures of lapachol and excipients obtained
in dynamic nitrogen atmosphere (50 ml min
1
) and at a heating rate of
10

Cmin
1
.
onset of 138.5

C (H=105.6 J g
1
). After this event, thermal
decomposition of material was observed at a temperature of
approximately 141

C (Fig. 1). The thermal behavior of Lpch

using DSC photovisual can be seen in Fig. 2. TG/DTG curves
indicate that the thermal decomposition process of Lpch occurs
in one stage of mass loss in the temperature range of 141260

C.
The DSC curves of the isolated solid excipients and binary
mixtures are presented in Figs. 3 and 4. The melting endotherm
Fig. 5. DSC photovisual of binary mixture of lapachol and excipients obtained in dynamic nitrogen atmosphere (50 ml min
1
) and at a heating rate of 10

Cmin
1
.
4 A.M. Lira et al. / Thermochimica Acta 457 (2007) 16
Fig. 6. DSC curves of Lpch:methylparaben mixtures in ratios (w/w) of 100:0,
75:25, 50:50, 25:75, and 0:100.
Fig. 7. DSC curves of Lpchcetostearyl alcohol mixtures in ratios (w/w) of
100:0, 75:25, 50:50, 25:75, and 0:100.
of the drug was well preserved in the majority of cases. How-
ever, there were slight changes in the peak shape with little
broadening or shifting towards the lower temperature. The DSC
curves of the physical mixtures of Lpch and edetic acid, propy-
Fig. 8. DSC curves of Lpchglyceryl-monostearate mixtures in ratios (w/w) of
100:0, 75:25, 50:50, 25:75, and 0:100.
Fig. 9. DSC curves of lapachol and liquid excipients obtained in dynamic nitro-
gen atmosphere (50 ml min
1
) and at a heating rate of 10

Cmin
1
.
lparaben, carbopol and cetomacrogol 1000 can be considered
to be the superposition of the DSC curves of the two individ-
ual components. These results showthat physical interactions of
components did not occur within the mixture. Photovisual DSC
was used to conrm the results obtained by conventional DSC
(Fig. 5).
In the thermal curve of the binary mixture of
Lpch/methylparaben, there was an appreciable downward
shift of the drug peak temperature, which can be indicative of
some drug-excipient solid interaction. DSC curves showed that
the peak at around 139 and 124

C, which was observed for Lpch

and methyparaben, respectively, disappeared in the eutectic
mixture (Fig. 4). However, a new peak at around 104

C, which
was not observed for Lpch and methyparaben, appeared in
the eutectic mixture. Binary mixtures of Lapch/methylparaben
in ratios (w/w) of 100/0, 75/25, 50/50, 25/75, and 0/100,
respectively, were prepared for thermal analysis (Fig. 6). Scott
et al. [16] used DSC to study the eutectic formation by a model
non-stereoidal anti-inammatory drug with a range of seven
terpene transdermal permeation enhancer. They found that DSC
studies on the ibuprofen:thymol mixtures indicated that they
Fig. 10. DSC curves of binary mixtures of lapachol and excipients obtained
in dynamic nitrogen atmosphere (50 ml min
1
) and at a heating rate of
10

Cmin
1
.
A.M. Lira et al. / Thermochimica Acta 457 (2007) 16 5
Fig. 11. DSC photovisual of binary mixture of lapachol and liquid excipients obtained in dynamic nitrogen atmosphere (50 ml min
1
) and at a heating rate of
10

Cmin
1
.
provide an example of a simple binary eutectic system with no
evidence of solid solution formation. A similar behavior was
observed in this study for the Lpch:methylparaben mixture.
In the DSC curves of the Lpchcetostearyl alcohol and
Lpchglyceryl-monostearate binary mixtures, the endothermic
peak of drug was broadened and shifted to a lower temper-
ature (Fig. 4). This occurred because cetostearyl alcohol and
glyceryl-monostearate showed a sharp endothermic peak at 53
and 60

C, respectively, corresponding to melting. Instead, DSC

photovisual showed that this effect was mainly due to the partial
dissolution of drug in melted excipients (Fig. 5). The disap-
pearance of the melting peak of drug is indicative of a strong
interaction, but not necessarily corresponding to incompatibil-
ity. The modication in drug thermal behavior was somewhat
intense, depending on the different degree of drug solubil-
ity in melted component and, for the same excipients, in the
mechanical treatment sustained by the sample [17]. In fact, a
similar effect was observed for other drugs, such as naproxen
[18], piroxicam [19], ketoprofen in mixtures with various PEGs
and was attributed to drug dissolution in the melted polymer
[20].
Figs. 7 and 8 show the DSC curves of the binary mixtures
of Lpch/cetostearyl alcohol and Lpch/glyceryl monostearate in
different proportions. As one can see, the DSC scans showed
small melting peaks even with such low drug content. These
interactions seemed to be very effective in physical mixtures
containing 50 and 25 wt% Lpch, and its melting point was not
possible to be detected. By applying DSC to look at interaction
between felodipine and excipients, Bikiaris et al. [21] explored
the physical state of a variety of new solid dispersion of hes-
peretin or felodipine. In the case of dispersions in PEG as well
as the physical mixtures, DSC, though fast rates were used, could
not detect the presence of crystalline drug. This was not because
the drug was amorphous, but because of the increased solubility
of the drugs in the liquid PEG at elevated temperatures.
Fig. 9 presents the DSCcurves of mineral oil, isodecyl oleate,
trietanolamine and glycerine. It was noted that the excipients
were thermally stable up to approximately 150

C. After this
temperature, processes of volatization and thermal decomposi-
tion of these materials occur. The thermal behavior of physical
mixtures of lapachol and excipients correspond to the superposi-
tion of the DSC curves of the two individual components. Lpch
demonstrated compatibility with all the liquid excipients stud-
ied, as shown in Fig. 10. DSC Photovisual analysis showed that
the viscosity of liquid excipients decreases with temperature,
therefore, their volume increases (Fig. 11).
4. Conclusion
Thermoanalytical methods were used extensively to evaluate
the physical properties of drugs, including melting and vaporiza-
tion temperatures and with the corresponding enthalpies, glass
transitions, vapor pressures, as well as to study the compatibility
and stability of the components of pharmaceutical prepara-
tions. Possible incompatibilities between active components and
different excipients were observed by using DSC. However, dif-
ferences in the DSC curves of binary mixtures and individual
compounds do not necessarily correspond to incompatibility.
For further conrmation of a substantial incompatibility, it is
important to associate with other analytical techniques. Con-
ventional and photovisual DSC curves of Lpch and excipients
showed interactions with glyceryl monostearate, methylparaben
and cetostearyl alcohol.
Acknowledgements
The authors acknowledge the nancial support by Conselho
Nacional de Desenvolvimento Cientco e Tecnol ogico (CNPq)
and Coordenac ao de Aperfeicoamento de Pessoal de Ensino
Superior (CAPES).
6 A.M. Lira et al. / Thermochimica Acta 457 (2007) 16
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